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DISSERTATION 


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Reprinted  from  THE  AMERICAN  JOURNAL  OF  PHYSIOLOGY     > 
Vol.  59,  No.  1,  February,  1922 


SOME  PLANT  SOURCES  OF  VITAMINS  B  AND  C1 

FRANCISCO  0.  SANTOS2 

From  the  Sheffield  Laboratory  of  Physiological  Chemistry,   Yale  University, 

New  Haven,  Conn. 

Received  for  publication  September  28,  1921 

In  the  problem  of  food  selection,  instinct  is  considered  by  many  per- 
sons to  be  a  fairly  safe  guide.  Therefore  it  is  often  assumed  that  the 
question  of  vitamin  deficiency  in  the  diet  needs  no  special  considera- 
tion. This,  however,  is  not  invariably  the  case.  In  the  Philippines, 
where  fruits  and  vegetables  abound  all  the  year  round,  deficiency  dis- 
eases have  occurred  and  are  occurring  among  the  Filipinos  as  well  as 
among  the  people  living  in  the  temperate  zones.  That  the  people  who 
can  obtain  the  foods  which  are  generally  considered  as  excellent  sources 
of  vitamins  should  be  subject  to  deficiency  diseases  may  be  due  to  one 
or  both  of  the  following  reasons:  They  are  either  eating  more  and  more 
of  artificially  treated  foods  or  are  not  eating  the  right  kind.  So  far  no 
systematic  investigation  of  the  vitamin  content  of  Philippine  food 
materials  has  been  made,  except  that  of  Brill  and  Alincastre  on  "the 
possible  maximum  vitamin  content  of  some  Philippine  vegetables"  (3). 
In  the  light  of  our  newer  knowledge  concerning  these  mysterious  sub- 
stances, the  above  work  appears  of  no  value.  Therefore  in  a  series  of 
studies  of  the  nutrition  of  the  Filipinos  it  became  highly  .desirable  that 
vitamin  tests  be  made  on  the  fruits  and  vegetables  which  are  being  or 
may  be  eaten  by  them. 

Several  vegetables  and  fruits  have  already  been  tested  for  vitamin 
content  by  different  investigators  (27),  (28),  (7),  (34),  (9),  (16),  (21), 
(30),  (31),  (32),  (33),  (17),  (18),  (26),  (24),  (11),  (4),  (8),  (20),  (35). 
A  thorough  review  of  the  subject  shows,  however,  that  there  are  still 
many  uninvestigated,  which  are  commonly  eaten.  The  present  work 
is  intended  as  a  beginning  of  a  series  of  tests  which  will  be  undertaken 
soon.  It  is  regretted  that  the  distance  from  the  Philippines  has  made 

1  The  data  in  this  paper  are  taken  from  the  dissertation  presented  by  the 
writer  for  the  degree  of  Doctor  of  Philosophy,  Yale  University,  1922. 

2  Traveling  Fellow  of  the  University  of  the  Philippines. 

310 


PLANT   SOURCES   OF   VITAMINS   B   AND    C  311 

the  selection  of  materials  limited  to  what  could  be  purchased  here  in 
the  States  and  what  were  furnished  by  Doctor  Kellogg  of  the  Battle 
Creek  Sanatorium,  the  United  Fruit  Co.,  and  Mr.  Ames,  Director  of  the 
Botanic  Garden  of  Harvard  University  in  Cuba,  to  whom  I  desire  to 
express  my  hearty  appreciation.  The  present  tests  became  dependent 
on  the  supply  of  materials  available. 

I.   TEST    FOR   VITAMIN    B 

Method.  In  order  to  obtain  a  comparative  estimate  of  the  vitamin 
content  of  the  materials  under  investigation  a  plan  similar  to  the  one 
used  by  Osborne  and  Mendel  (32)  has  been  adopted.  The  fruits  and 
vegetables  were  dried  in  an  air  current  at  less  than  85°C.  The  dried 
materials  were  ground  finely  and  then  made  into  pills  of  1  gram, 
|  gram,  or  less,  according  to  the  need  of  the  experiments.  Glucose  syrup 
was  used  as  adhesive. 

Osborne  and  Mendel  (31)  have  prepared  a  diet  for  albino  rats  which 
has  been  shown  to  be  complete  in  every  respect  except  that  it  lacks 
vitamin  B.  '  This  mixture  was  slightly  modified  in  percentage  com- 
position as  follows: 

Casein 18 

Salt  mixture3 4 

Starch 50 

Butter  fat 9 

Lard 19 

On  this  food  the  rats  slowly  lose  appetite  and  after  a  time  decline  in 
weight.  Then  if  the  diet  is  not  changed  or  improved  by  the  addition  of 
substances  containing  vitamin  B,  death  ensues.  In  most  cases  the 
animals  promptly  improve  in  appetite  when  vitamin  B  is  given.  But 
when  the  source  of  vitamin  is  too  bulky  so  that  it  prevents  the  ingestion 
of  sufficient  of  the  standard  food,  the  total  intake  of  all  essentials  may 
become  inadequate.  The  ability  to  cause  the  recovery  of  rats  which 
have  been  declining  in  weight  on  a  diet  which  is  known  to  be  complete 
in  every  respect  except  that  it  lacks  vitamin  B,  has  been  taken  as  a 
proof  that  the  material  in  question  contains  the  latter. 

The  feeding  technique  was  essentially  like  that  commonly  employed 
in  this  laboratory  and  described  by  Ferry  (12).  Protocols  of  body 
weights  and  food  intakes  are  given  in  tables,  and  some  of  the  data 

8  The  salt  mixture  used  is  that  described  by  Osborne  and  Mendel,  Journ. 
Biol.  Chem.,  1917,  xxxii,  317. 


A78G55 


312 


FRANCISCO   O.   SANTOS 


are  also  presented  in  graphic  forms  in  charts.     (Tables  1  to  10  and 
charts  1  to  12.) 

TABLE  1 

Mongo;  body  weight  and  weekly  intake  of  standard  diet 


FEEDING 
PERIOD 

ONE   GRAM  SUPPLEMENT 

Rat  3d" 

Ratio  9 

Rat  9  9 

Rat  7  9 

Body 
weight 

Diet 

Body 
weight 

Diet 

Body 
weight 

Diet 

Body 
weight 

Diet 

weeks 

grams 

grams 

grams 

grams 

grams 

grams 

grams 

grams 

150 

168 

66 

178 

83 

64 

70 

194 

95 

94 

93 

1 

174 

? 

94 

33 

94 

43 

99 

46 

2 

164 

? 

90 

34 

97 

30 

92 

29 

3 

152x 

? 

85 

16 

90 

20 

82 

20 

4 

166 

46 

75 

16 

84 

16 

81 

16 

5 

178 

44 

64x 

j 

80x 

12 

76 

16 

6 

190 

46 

85 

29 

98 

32 

76x 

12 

7 

210 

53 

102 

36 

108 

36 

97 

36 

8 

220 

104 

36 

107 

32 

115 

45 

9 

117 

46 

115 

38 

122 

46 

10 

126 

40 

132 

44 

FEEDING  PERIOD 

ONE-HALF  GRAM  SUPPLEMENT 

Rat3cf 

Rat  11  9 

Rat  9  9 

Body  weight 

Diet 

Body  weight 

Diet 

Body  weight 

Diet 

weeks 

grams 

grams 

grams 

grams 

grams 

grams 

220 

218 

166 

115 

1 

202 

? 

153 

34 

122 

40 

2 

195 

? 

150 

29 

123 

40 

3 

184x 

? 

140 

25 

117 

27 

4 

192 

45 

130x 

23 

lOOx 

? 

5 

198 

50 

143 

34 

118 

36 

6 

198 

43 

152 

37 

120 

36 

7 

196y 

54 

154 

52 

132 

40 

8 

206 

56 

154y 

42 

136 

37 

9 

231 

55 

172 

50 

DISCUSSIONS    OF    THE    MATERIALS    USED    AND    THE    FEEDING    EXPERI- 
MENTS.    MONGO:  (Phaseolus  mongo  L.).     This   bean    contains   about 


PLANT  SOURCES  OF  VITAMINS  B  AND   C 


313 


20  per  cent  protein  (N  X  6.25)  (2).     It  is  extensively  used  by  the  Fil- 
ipinos  and  is  practically  the  only  food  of  the  people  afflicted  with 

TABLE  2 
Togi;  body  weight  and  weekly  intake  of  standard  diet 


ONE  GRAM  SUPPLEMENT 

rXIDING  PERIOD 

Rat  1  9 

Rat8  <f 

Rat  2  9 

Body  weight 

Diet 

Body  weight 

Diet 

Body  weight 

Diet 

weeks 

grams 

grams 

grams 

grams 

grams 

grams 

136 

134 

144 

67 

144 

150 

95 

154 

1 

140 

? 

112 

41 

138 

? 

2 

116x 

? 

103 

29 

102x 

? 

3 

136 

43 

98x 

? 

136 

56 

4 

150 

49 

118 

35 

160 

62 

5 

152 

52 

135 

31 

162 

50 

6 

153 

36 

149 

42 

174 

46 

7 

156 

40 

164 

35 

180 

8 

162 

47 

175 

36 

ONE-HALF  GRAM  SUPPLEMENT 

Rat  11  ? 

Rat  13  c? 

Rat6  9 

130 

144 

74 

156 

165 

98 

' 

1 

156 

33 

162 

60 

105 

40 

2 

154 

35 

159 

46 

100 

34 

3 

141 

16 

150 

26 

92 

24 

4 

130 

16 

140 

17 

92 

28 

5 

118x 

15 

128 

? 

94 

27 

6 

130 

44 

122x 

? 

98 

23 

7 

146 

50 

132 

50 

92 

21 

8 

153 

36 

144 

39 

90 

23 

9 

159 

39 

153 

42 

82x 

24 

10 

166 

38 

165 

46 

98 

41 

11 

177 

47 

120 

46 

12 

135 

39 

13 

148 

48 

wet  beriberi  who  at  the  same  time  cannot  buy  milk    The  mongo  used 
was  bought  from  a  Chinese  grocery  store  at  New  Haven.    The  bean 


314 


FRANCISCO   O.   SANTOS 


TABLE  3 
Sweet  potato  leaves;  body  weight  and  weekly  intake  of  standard  diet 


FEEOINO 
PERIOD 

ONE    GRAM   SUPPLEMENT 

ONE-HALF  GRAM  SUPPLEMENT 

Ratl  9 

Rat  2  9 

Rat  3d" 

Rat  4  d" 

Body 

weight 

Diet 

Body 

weight 

Diet 

Body 
weight 

Diet 

Body 
weight 

Diet 

weeks 

grams 

grams 

grams 

grams 

grams 

grams 

grams 

grams 

132 

126 

198 

186 

1 

128 

43 

124 

40 

190 

47 

181 

43 

2 

128 

37 

113 

24 

184 

49 

170 

39 

3 

113x 

15 

lOOx 

15 

168 

41 

159 

29 

4 

124 

35 

115 

27 

148x 

15 

138x 

16 

5 

136 

49 

134 

44 

150 

29 

128 

21 

6 

144 

49 

144 

48 

168 

52 

121 

18 

7 

150 

44 

154 

39 

178 

44 

132 

36 

8 

194 

49 

140 

29 

9 

143z 

37 

10 

159 

52 

11 

153y 

48 

12 

164 

50 

13 

180 

45 

14 

195 

49 

z  =  0.2  gram  yeast  was  given  as  daily  supplement. 

TABLE  4 
Bilimbi;  body  weight  and  weekly  intake  of  standard  diet 


ONE    GRAM   SUPPLEMENT 


FEEDING  PERIOD 

Rat  26  9 

Rat6  $ 

Body  weight 

Diet 

Body  weight 

Diet 

weeks 

grams 

grams 

grams 

grams 

127 

140 

1 

.130 

37 

136 

31 

2 

132 

38 

130 

27 

3 

137 

41 

124 

27 

4 

130 

33 

122 

? 

5 

116x 

28 

120x 

? 

6 

HOy 

11 

115d 

? 

7 

137 

52 

132 

41 

8 

160 

67 

d  =  given  one  gram  duhat  daily  instead  of  bilimbi. 


PLANT   SOURCES   OF   VITAMINS   B   AND    C 


315 


TABLE  5 
Duhat;  body  weight  and  weekly  intake  of  standard  diet 


FEEDING  PERIOD 

ONE   GRAM  SUPPLEMENT 

Rat  18  cf 

Rat6  9 

Body  weight 

Diet 

Body  weight 

Diet 

weeks 

grams 

grams 

grams 

grams 

150 

140 

1 

160 

54 

136 

31 

2 

154 

46 

130 

27 

3 

154 

36 

124 

27 

4 

153 

? 

122 

j 

5 

143 

? 

120 

? 

6 

142 

? 

115x 

? 

7 

140 

? 

132 

41 

8 

138 

? 

9 

128x 

? 

10 

144 

35 

11 

152 

55 

12 

160 

46 

13 

176 

50 

The  supply  of  duhat  was  exhausted 

TABLE  6 
Okra;  body  weight  and  weekly  intake  of  standard  diet 


ONE  GRAM  SUPPLEMENT 

ONE-HALF  GRAM  SUPPLEMENT 

FEEDING  PERIOD 

Ratl  9 

Rat  13  cf 

Rat  4  d" 

Body  weight 

Diet 

Body  weight 

Diet 

Body  weight 

Diet 

weeks 

proms 

grams 

grams 

grams 

proms 

proms 

162 

177 

159 

159 

195 

1 

159 

44 

148 

38 

192 

47 

2 

138 

? 

129x 

? 

178 

45 

3 

118x 

? 

146 

45 

178 

62 

4 

142 

53 

156 

48 

166 

32 

5 

160 

60 

167 

45 

152x 

29 

6 

164 

43 

174 

50 

170 

47 

7 

179 

47 

180 

52 

180 

40 

8 

178 

48 

9 

186 

51 

10 

204 

59 

316  FRANCISCO   O.   SANTOS 

TABLE  7 
Artichokes;  body  weight  and  weekly  intake  of  standard  diet 


ONE  GRAM  SUPPLEMENT 

FEEDING  PERIOD 

Rat  23  d" 

Rat  24  tf 

Rat  22  <? 

Body  weight 

Diet 

Body  weight 

Diet 

Body  weight 

Diet 

weeks 

grams 

grams 

grams 

grams 

grams 

grams 

175 

226 

205 

180 

224 

198 

1 

186 

62 

234 

51 

184 

36 

2 

191 

48 

220 

39 

168x 

22 

3 

182 

44 

212 

28 

181 

45 

4 

169 

? 

195 

31 

204 

62 

5 

161 

36 

184x 

22 

218 

64 

6 

155x 

23 

197 

35 

222 

7 

177 

50 

201 

36 

8 

188 

46 

204 

45 

9 

190 

54 

218 

50 

10 

190y 

48 

210y 

32 

11 

208 

59 

240 

56 

FEEDING  PERIOD 

ONE-HALF  GRAM  SUPPLEMENT 

Rat  15  rf1 

Rat  25  d" 

Body  weight 

Diet 

Body  weight 

Diet 

weeks 

grams 

grams 

grams 

grams 

212 

212 

218 

1 

196 

34 

220 

70 

2 

178x 

19 

224 

49 

3 

181 

? 

218 

38 

4 

191 

44 

209 

34 

5 

196 

30 

195 

36 

6 

196y 

? 

190 

35 

7 

220 

60 

178x 

31 

8 

185 

50 

9 

185 

48 

10 

192 

40 

11 

190y 

? 

12 

204 

53 

13 

220 

57 

14 

224 

PLANT   SOUKCES   OF  VITAMINS   B   AND    C 

TABLE  8 
Avocado;  body  weight  and  weekly  intake  of  standard  diet 


317 


FEEDING 
PERIOD 

ONE    GRAM  SUPPLEMENT 

ONE-HALF  GRAM  SUPPLEMENT 

Rat  7  9 

Rat  21  o* 

Rat  17  o" 

Rat  20  cf 

weight           Diet 

wligtt          Diet 

Body            n-  t 
weight 

Body 
weight 

Diet 

weeks 

grams          grams 

grams          grams 

grams          grams 

grams 

grams 

141 

180 

148 

148 

132 

187 

156 

154 

1 

121            26 

194            49 

160            56 

153 

45 

2 

112x            ? 

192            48 

158            48 

147 

42 

3 

139            44 

178            36 

147            26 

138 

26 

4 

150            56 

155x          18 

130x            ? 

120x 

20 

5 

150            42 

207            69 

160            53 

141 

42 

6 

158            54 

240            71 

178            59 

152 

49 

7 

240            61 

192            54 

167 

50 

8 

244            58 

200            48 

172 

47 

FEEDING  PERIOD 

ONE  -TENTH  AND  FOUB-TENTH  GRAM  SUPPLEMENTS 

Rat  16  o" 

Rat  19  d" 

Body  weight 

Diet 

Body  weight 

Diet 

weeks 

grams 

grams 

grams 

grams 

202 

156 

154 

1 

172    . 

26 

160 

51 

2 

168 

31 

154 

37 

3 

178 

44 

154 

33 

4 

176 

39 

148 

27 

5 

178 

41 

142 

28 

6 

170x 

45 

134x 

25 

7 

162 

38 

130 

30 

8 

154xx 

28 

128xx 

37 

9 

165 

34 

142 

36 

10 

160 

48 

153 

43 

11 

170 

47 

160 

58 

12 

162y 

35 

160y 

51 

13 

176 

64 

184 

59 

14 

200 

72 

15 

206 

x  =  0.1  gram. 

xx  =  0.4  gram. 

318 


FRANCISCO   O.   SANTOS 


was  soaked  in  water  for  a  day,  then  cooked  as  is  done  for  culinary  use. 
The  mashed  mongo  including  the  water  in  which  it  was  boiled,  was 
then  dried,  powdered  and  finally  made  into  pills.  Rats  declining  in 
weight  easily  recovered  their  original  weight  under  a  daily  supplement 
of  1  gram  to  the  standard  ration  free  from  vitamin  B.  With  %  gram, 
one  animal  recovered  its  original  weight,  while  two  did  not  in  about  the 
same  length  of  time  thus  indicating  that  the  latter  dose  is  about  the 
minimum  for  this  species. 

TOGI:  This  is  the  name  given  to  the  sprouted  mongo.  The  bean 
was  first  soaked  in  water  for  24  hours,  then  allowed  to  sprout  in  the 
dark  for  2  days,  cooked,  dried  including  the  water  used  for  boiling,  and 

TABLE  9 
Banana  flower  bud;  body  weight  and  weekly  intake  of  standard  diet 


ONE   GRAM  SUPPLEMENT 

FEEDING  PERIOD 

Rat  27  9 

Rat  28  9 

Rat  10  9 

Body  weight 

Diet 

Body  weight 

Diet 

Body  weight 

Diet 

weeks 

grams 

grams 

grams 

grams 

grams 

grams 

150 

184 

126 

1 

155 

40 

186 

51 

125 

32 

2 

152 

38 

185 

43 

122 

30 

3 

154 

45 

188 

51 

119 

28 

4 

147 

30 

178 

34 

115 

27 

5 

132x 

? 

161x 

26 

110 

28 

6 

H5y 

? 

145y 

27 

110 

28 

7 

145 

60 

178 

67 

108 

26 

8 

162 

55 

196 

62 

96x 

20 

Died 

powdered.  Both  1  gram  and  ^  gram  daily  supplements  caused  quick 
recovery  in  weight  of  rats  which  had  been  declining  on  a  diet  deficient 
in  vitamin  B.  One-half  gram  of  the  togi  was  more  potent  than  \  gram 
of  the  mongo.  This  indicates  that  the  vitamin  B  in  the  mongo  is  in- 
creased in  sprouting;  a  fact  contrary  to  the  finding  of  Grijns  (cited  by 
Chick  and  Hume  (6)  )  that  the  anti-beriberi  vitamin  is  lessened  in 
amount  as  germination  takes  place.  Togi  then  has  three  advantages 
over  mongo:  greater  digestibility  (1)  and  greater  content  of  vitamins 
B  and  C  (see  later). 

SWEET  POTATO  LEAVES:  (Batatas  batatas}.    The  leaves  and  young 
shoots  of  sweet  potato  are  used  as  vegetables  in  making  stews  and  simi- 


PLANT   SOURCES   OF   VITAMINS   B   AND    C 


319 


lar  dishes.  The  material  for  study  was  obtained  dry.  One  gram  daily 
supplement  fed  to  rats  which  had  been  declining  in  weight  on  a  diet 
deficient  in  vitamin  B  caused  immediate  recovery.  In  about  the  same 
length  of  time  \  gram  caused  only  partial  recovery.  The  latter  dosage 
is  not  as  efficient  as  -~  gram  of  dried  brewery  yeast.4 

TABLE  10 
Bamboo  shoots;  body  weight  and  weekly  intake  of  standard  diet 


FEEDING  PERIOD 

OXE  GRAM  SUPPLEMENT 

Rat  12  <? 

Rat  15  cf 

Rat  16  d* 

Body  weight 

Diet 

Body  weight 

Diet 

Body  weight 

Diet 

weeks 

grams 

grams 

grams 

grams 

grams 

grams 

175 

203 

164 

202 

1 

205 

64 

178  . 

63 

201 

74 

2 

204 

65 

185 

55 

202 

67 

3 

188 

42 

176 

37 

176 

26 

4 

176 

19 

158x 

? 

158x 

13 

5 

157x 

10 

162 

26 

168 

44 

6 

172 

36 

163 

41 

173 

48 

7 

191 

31 

172 

38 

180 

41 

s 

195 

46 

182 

38 

172 

26 

£ 

196d 

43 

ISSxx 

42 

168xx 

31 

10 

192 

42 

190 

41 

178 

44 

11 

170xx 

22 

196 

38 

176 

39 

12 

184 

44 

208 

64 

178 

41 

13 

198 

63 

212 

54 

170y 

45 

14 

198 

55 

15 

198y 

49 

16 

210 

60 

17 

222 

68 

206 

x  =  1  gram. 

xx  =  2  grams. 

d  =  diet  only  again. 

BILIMBI:  (Averrhoa  carambola  L.).  Only  a  small  sample  in  dry 
condition  was  obtained,  so  only  the  one  gram  supplement  could  be 
tested.  After  7  days  of  1  gram  daily  supplement,  no  improvement  in 
the  weight  of  the  animals  was  found.  The  intake  of  basal  diet  did  not 

4  The  dried  brewery  yeast  used  was  kindly  furnished  by  Osborne  and  Mendel 
from  their  stock. 


320 


FRANCISCO   O.    SANTOS 


PLANT   SOURCES   OF  VITAMINS   B   AND    C 


321 


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322 


FRANCISCO    O.    SANTOS 


CHART  /.  ARTICHCKKS  AS  SOURCE 
OF  TOMlX  B. 


PLANT   SOUKCES   OF  VITAMINS   B   AND    C 


323 


CHART  3-  BAKANA  FLOWER  BUD 
AS  SOORCE  OF  VITAMIM  B. 


324 


FRANCISCO   O.    SANTOS 


increase.  But  when  -±  gram  yeast  was  given  to  rat  26,  instead  of  bi- 
limbi,  it  gained  weight  very  rapidly.  Duhat  improved  the  weight  and 
appetite  of  rat  6. 

DUHAT:  (Eugenia  jambolana  L.).  Only  enough  material  for  test- 
ing on  one  animal  was  obtained.  One  gram  daily  supplement  caused 
the  recovery  of  rat  18  which  had  been  declining  in  weight  on  a  diet 


CHART  iO.  BAMBOO  SHOOTS  AS 
SOURCE  OF  YTTAHIH  B. 


deficient  in  vitamin  B.    The  incomplete  experiment  with  rat  6  also 
shows  that  duhat  caused  an  improvement  in  appetite  and  in  weight. 

OKRA:  (Abelmoschus  esculentus).  Although  this  is  not  used  in  the 
Philippines,  it  is  a  tropical  plant  which  can  be  introduced.  The  fruit 
was  boiled,  dried  including  the  water  in  which  it  was  boiled,  and  then 
powdered.  Both  1  gram  and  |  gram  daily  supplements  caused  the 
recovery  in  weight  of  rats  declining  due  to  lack  of  vitamin  B. 


PLANT  SOURCES  OF  VITAMINS  B  AND   C 


[325 


326 


FRANCISCO   O.   SANTOS 


AHTICHOKES:  (Cynara  scolymus).  Though  this  has  little  nutritive 
value  it  is  eaten  as  a  delicacy.  Its  use  in  the  Philippines  is  not  known 
but  may  be  introduced.  The  edible  portion  was  dried  and  powdered. 
The  behavior  of  rats  22,  23,  24  shows  that  1  gram  daily  supplement  is 
probably  just  enough  to  cause  recovery  in  weight  of  rats  which  had  been 
declining  due  to  lack  of  vitamin  B.  One-half  gram  daily  was  found  to 
be  insufficient. 

AVOCADO:  (Per  sea  per  sea).  This  fruit  is  rich  in  fat  (25),  and  is 
being  increasingly  used  not  only  in  the  tropics  but  also  in  the  temper- 


CHART  12.   CURVES  SHOWING  COMPARATIVE  RECOVER*  OF  GROWTH 
WITH  VITAMIX  B  FROM  THE  JHFPEREHT  KAiTEJUALS  INVESTIGATED. 
ONE  HAJ.F  GUAM  PAILY  SUPPLBHEXT. 


ate  zones.  The  edible  portion  of  the  dried  fruit  was  ground  and  ex- 
tracted with  ether  to  remove  most  of  the  fat.  Rats  which  had  been 
declining  in  weight  due  to  lack  of  vitamin  B,  easily  recovered  on  the 
administration  of  either  1  gram  or  |  gram  extracted  avocado  daily  in 
addition  to  the  standard  ration.  One-tenth  gram  was  found  to  be 
insufficient;  and  ^  gram  seems  to  be  just  enough  to  cause  maintenance 
but  not  complete  recovery  of  weight. 

BANANA  FLOWER  BUD:  (Musa  sapientum).  This  is  used  in  making 
salads.  The  sample  was  obtained  in  an  already  dried  condition  and 
the  variety  was  undetermined.  In  the  Philippines  only  the  buds  of 


PLANT   SOURCES   OF   VITAMINS   B   AND   C  327 

certain  varieties  are  used,  because  some  have  bitter  taste.  Even  a 
1-gram  daily  supplement  was  not  sufficient  to  check  the  decline  in  weight 
of  rats  which  had  been  taking  vitamin  B  free  diet.  Similar  results  were 
obtained  with  mice,  which  could  not  even  be  maintained  by  daily 
addition  of  \  gram  of  banana  flower  bud  to  the  basal  diet. 

BAMBOO  SHOOTS:  (Bambusa  sp.).  Canned  material  from  Japan  was 
used.  The  shoots  including  the  water  in  which  they  were  boiled  were 
dried  and  powdered.  Rat  15  after  5  weeks  of  1  gram  daily  supple- 
ment to  the  basal  diet  recovered  its  weight,  and  when  in  addition  to 
this  supplement  alcoholic  extract  of  the  powdered  shoots  was  incorpor- 
ated with  the  basal  diet  the  animal  continued  gaining  in  weight.  Enough 
of  the  extract  was  added  to  the  basal  diet  so  that  an  amount  corre- 
sponding to  approximately  1  gram  of  the  original  shoot  was  eaten  by  the 
rat  daily  in  addition  to  the  1  gram  daily  supplement  given  separately. 
The  alcoholic  extract  was  first  offered  separately  also  but  was  refused 
by  the  animal.  Rat  16  after  5  weeks  of  1  gram  daily  supplement  did 
not  recover  its  original  weight.  Then  a  2-gram  daily  supplement  was 
given.  Neither  weight  nor  appetite  improved.  Oftentimes  the  rat 
was  observed  to  have  eaten  only  the  bamboo  supplement  and  very 
little  of  the  standard  food.  This  is  a  case  in  which  the  limited  capacity 
of  the  animal  prevents  it  from  eating  more  of  the  necessary  basal  diet 
when  the  bulky  food  containing  the  vitamin  was  preferred.  It  finally 
recovered  its  weight  on  the  administration  of  -^  gram  yeast  daily. 
Rat  12  did  not  fully  recover  its  weight  after  about  the  same  length  of 
time.  To  determine  whether  there  is  really  vitamin  B  in  the  bamboo 
shoots,  the  rat  was  placed  on  the  basal  diet  again.  Weight  and  ap- 
petite both  declined.  Then  2-grams  daily  supplement  of  bamboo  was 
added  to  the  basal  ration.  A  result  similar  to  that  with  rat  16  was 
obtained. 

One  gram  daily  was  also  given  to  mice  which  had  been  declining  in 
weight  due  to  lack  of  vitamin  B  in  the  diet,  but  no  improvement  was 
noticed. 

On  the  assumption  that  the  vitamin  B  and  the  antineuritic  vitamin 
are  related,  an  experiment  was  tried  with  pigeons.  Three  pigeons 
(nos.  1,  2  and  3)  were  stuffed  with  polished  rice,  equal  in  weight  to 
from  ^  to  Y^  of  their  body  weight,  plus  6  grams  of  powdered  bamboo 
shoots  daily.  All  died  of  atrophic  polyneuritis  (13),  pigeons  1  and 
2  after  18  days,  and  pigeon  3  after  35  days.  Of  the  three  controls  with- 
out bamboo,  pigeon  4  died  of  polyneuritis  after  12  days;  pigeon  5  on 
the  19th  day  showed  a  tendency  to  fall  on  one  side  (a  sign  of  polyneu- 

THB  AMERICAN  JOURNAL  OP  PHYSIOLOGY,    VOL.   59,   NO.    1 


328  FRANCISCO   O.  SANTOS 

ritis)  and  was  cured  by  the  injection  of  an  alcoholic  extract  from  6 
grams  of  wheat  embryo.  Control  pigeon  6  died  of  polyneuritis  after 
31  days.  Pigeons  3  and  6  always  vomited  the  food  that  was  stuffed 
in  them  except  during  the  4  or  5  days  before  their  death.  It  thus 
appears  that  bamboo  shoots  have  little  vitamin  B. 

TAMARIND:  (Tamarindus  indicus).  The  young  leaves  are  used  as 
vegetables  and  give  sour  taste  to  the  broth.  The  rats  could  not  be 
made  to  eat  the  material.  Even  incorporating  the  powdered  leaves 
with  the  basal  diet  was  unsuccessful.  Neutralization  of  the  sour  taste 
with  sugar  was  tried  but  too  much  sugar  was  required  and  thus  the 
material  became  bulky.  The  alcoholic  extract  was  also  refused  by  the 
rats. 

PIGWEED:  (Cycloloma  platyphyllwri) .  This  is  also  used  in  making 
stews  and  gives  bitter  taste  to  the  broth.  Like  the  tamarind,  it  was 
distasteful  to  the  rats. 

II.   TEST  FOR  VITAMIN   C 

Scurvy,  like  beriberi,  is  found  in  the  Philippines,  being  especially 
common  among  children  under  2  years  of  age.  Poor  nutrition  is  the 
cause  of  this.  Milk  is  only  within  the  reach  of  a  few.  Cases  are  com- 
mon in  which  mothers  who  have  poor  or  little  milk  give  the  broth  of 
boiled  polished  rice  to  the  children  as  a  substitute  for  milk.  This  milk 
substitute  is  of  course  poor  in  both  vitamins  B  and  C.  Since  vitamin 
C  is  destroyed  or  diminished  in  potency  under  the  influence  of  heat, 
as  in  cooking  and  drying  (22),  only  those  materials  which  promised 
useful  results  were  tested.  The  protocols  of  body  weights  are  given  in 
graphic  form  in  the  appendix  (charts  13  and  14). 

Method:  Guinea  pigs  were  used  as  the  experimental  animals.  Hess 
and  Unger's  (23)  scorbutic  diet  of  hay,  oats,  plus  0.5  to  1  cc.  of  cod 
liver  oil  daily  and  water  ad  libitum  was  used  as  the  basal  diet  in  most 
of  the  tests.  The  oil  was  given  by  a  pipette.  At  first  finely  chopped 
hard  boiled  egg  (yolk  plus  white)  was  offered  instead  of  the  cod  liver 
oil,  but  the  animals  would  not  eat  the  former.  In  the  experiment  with 
avocado  no  use  of  cod  liver  oil  was  made.  On  the  basal  ration  control 
guinea  pigs  (nos.  2,  13  and  14)  succumbed  to  scurvy  after  16,  26  and 
27'  days  respectively.  In  all  of  the  guinea  pigs  autopsied  it  was  found 
that  the  hemorrhages  were  more  severe  in  those  that  were  not  given 
oil  than  in  those  animals  that  ate  the  improved  diet.  At  first  curative 
experiments  were  tried;  but  it  was  difficult  to  make  the  guinea  pigs  eat 
when  they  were  already  suffering  from  scurvy  and  therefore  preventive 


PLANT   SOURCES   OF   VITAMINS   B   AND    C 


329 


tests  were  performed.  In  the  curative  experiments  the  animals  de- 
monstrated the  externals  signs  of  scurvy  such  as  the  scorbutic  position 
(22),  crying  when  handled,  painful  or  swollen  limbs,  and  pale  gums, 
before  the  materials  to  be  tested  were  given  as  supplement  to  the  scor- 
butic diet. 


DISCUSSIONS  OF  THE  MATERIALS  USED  AND  THE  FEEDING  EXPERI- 
MENTS. Inasmuch  as  in  every  day  nutrition  the  materials  to  be  tested 
here  are  never  eaten  alone,  since  the  amount  that  an  individual  can 
eat  in  a  day  is  dependent  in  part  on  his  alimentary  capacity,  and  since, 
further,  our  object  was  to  get  a  comparative  estimate  of  the  possible 
value  of  the  foods  as  practical  sources  of  vitamin  C,  a  daily  supple- 
ment of  10  grams  to  each  guinea  pig  in  addition  to  the  basal  ration  was 
considered  the  maximum. 


330 


FHANCISCO    O.    SANTOS 


MONGO:  (Phaseolus  mongo  L.).  In  all  cases  the  bean  was  ground 
and  soaked  in  water,  to  make  it  more  palatable,  before  being  offered  to 
the  animals. 

Ten  grams  daily  curative  supplement,  Guinea  pig  4  began  to  drag 
the  hind  legs  after  17  days  of  scorbutic  ration.  Mongo  was  now  given 


5  £rams  (toiled)  daily  supplement 
(preventive) 


separately  as  supplement.     The  decline  in  weight  continued  and  on  the 
33rd  day  it  died  of  scurvy. 

Guinea  pig  1  showed  external  symptoms  of  scurvy  on  the  14th  day 
of  scorbutic  diet.  When  mongo  was  given,  the  animal  improved  in 
appearance  and  recovered  the  original  weight.  On  the  89th  day  it 
was  chloroformed  and  autopsy  showed  little  hemorrhage  in  the  right 
hind  leg. 


PLANT   SOURCES   OF  VITAMINS   B   AND    C  331 

Ten  grams  daily  preventive  supplement.  Guinea  pig  15  never  ate 
all  of  the  mongo.  After  25  days,  the  animal  had  difficulty  in  moving 
the  hind  legs,  and  on  the  36th  day  it  died  of  scurvy. 

Guinea  pig  16  did  not  eat  all  of  the  mongo  for  the  first  2  weeks.  On 
the  57th  day  the  animal  began  dragging  the  hind  legs,  and  on  the  64th 
day  it  died.  Autopsy  showed  the  signs  of  scurvy. 

Five  grams  daily  preventive  supplement.  Guinea  pig  11  died  on  the  40th 
day  without  showing  external  sign  of  scurvy  except  pale  gums. 
Autopsy  showed  slight  hemorrhages  at  the  leg  joints. 

Guinea  pig  12  on  the  33rd  day  cried  when  the  left  hind  leg  was  touched. 
Soon  an  eye  became  blind  and  on  the  67th  day  it  died  of  scurvy. 

TOGI:  Fiirst  (14),  Chick  and  Hume  (6),  Cohen  and  Mendel  (7), 
Chick  and  Delf  (5)  and  others  have  shown  that  the  vitamin  C  of  dried 
peas  and  lentils  is  much  increased  in  germination.  Wiltshire  (36) 
has  successfully  used  germinated  haricot  beans  which  were  boiled  for 
10  minutes  in  the  treatment  of  human  scurvy.  Gerstenberger  (15) 
found  that  malt  soup  extract  prepared  from  malt  sprouted  in  a  cer- 
tain way  has  high  antiscorbutic  value.  Recently  it  has  been  shown 
that  oxidation  (10)  is  the  chief  factor  in  the  destruction  of  this  vita- 
min. Evidently  the  method  of  cooking  has  something  to  do  with  the 
destruction  of  vitamin  C.  Adkins  (1)  has  found  that  germinated  bean 
is  easier  to  digest  than  the  ungerminated.  In  the  light  of  the  above 
findings  togi  ought  to  be  of  great  value  in  nutrition.  It  is  cheap  and 
at  the  same  time  easily  obtainable.  In  anticipation  of  its  high  con- 
tent of  vitamin  C,  5  grams  daily  supplement  to  the  scorbutic  diet  was 
first  tried.  The  togi  used  was  weighed  and  given  to  the  animals  with- 
out the  outer  seed  coat. 

Five  grams  daily  curative  supplement.  Guinea  pig  3  having  devel- 
oped external  signs  of  scurvy  on  the  16th  day  of  scorbutic  diet  was  given 
fresh  togi  daily  as  supplement.  It  gradually  improved  in  appearance 
and  increased  in  weight  over  that  of  the  original.  On  the  89th  day  it 
was  chloroformed  and  no  sign  of  scurvy  was  shown  by  autopsy. 

Guinea  pig  5  showed  signs  of  scurvy  on  the  13th  day  of  scorbutic  diet, 
whereupon  it  was  given  fresh  togi  daily  as  supplement.  It  gradually 
recovered  the  original  weight.  On  the  91st  day  it  was  chloroformed. 
Autopsy  showed  nothing  abnormal. 

Guinea  pig  6  on  scorbutic  diet  developed  signs  of  scurvy  on  the  15th 
day  and  was  then  given  fresh  togi  as  supplement.  On  the  90th  day  it 
weighed  100  grams  over  the  initial  weight.  When  autopsied  after  being 
chloroformed  no  sign  of  scurvy  was  found. 


332  FRANCISCO   O.   SANTOS 

One  gram  daily  preventive  supplement.  Guinea  pig  9  was  given 
togi  as  supplement  in  addition  to  the  scorbutic  diet.  On  the  60th  day 
it  began  dragging  the  hind  legs,  and  on  the  78th  day  died  of  severe 
scurvy. 

Guinea  pig  10  was  treated  the  same  as  guinea  pig  9.  On  the  46th 
day  it  died  without  external  sign  of  scurvy.  Slight  hemorrhages  at 
the  limbs  were  seen  when  autopsied. 

Five  grams  boiled  (10  minutes')  togi  daily  preventive  supplement. 
Guinea  pig  7  died  of  scurvy  after  34  days  on  the  scorbutic  diet,  plus 
boiled  togi  daily.  Guinea  pig  8  on  the  same  food  as  guinea  pig  7  died 
of  scurvy  on  the  38th  day. 

To  both  animals  the  water  in  which  the  togi  was  boiled  was  also 
offered.  Care  was  taken  to  boil  the  togi  in  the  least  amount  of  water 
possible,  and  boiling  was  conducted  in  a  covered  vessel  as  is  done  in 
every  day  cooking. 

AVOCADO:  (Per sea  persea).  This  fruit  is  eaten  raw,  and  if  it  could  be 
shown  to  be  high  in  vitamin  C,  it  would  be  of  great  value.  Unfor- 
tunately the  guinea  pigs  would  not  eat  the  avocado;  so  both  the  pre- 
ventive and  curative  experiments  were  unsuccessful. 

SUMMARY 

Several  plant  foods  were  tested  for  their  content  of  vitamins  B  and  C. 

Togi,  okra  and  avocado  were  found  to  be  comparatively  high  in  vita- 
min B.  One-half  gram  of  each  of  them  as  daily  supplement  to 
the  standard  vitamin  B  free  diet  caused  the  recovery  in  weight  of 
rats  which  had  been  declining  because  of  lack  of  this  accessory  food 
factor. 

Mongo,  sweet  potato  leaves  and  duhat  contain  enough  vitamin  so  that 
1  gram  of  them  as  daily  supplement  caused  the  recovery  in  weight  of 
rats  which  had  been  declining  due  to  lack  of  vitamin  B. 

Artichokes,  bilimbi,  banana  flower  bud  and  bamboo  shoots  are  relatively 
poor  in  vitamin  B. 

The  vitamin  B  in  mongo  was  increased  in  germination,  a  fact  con- 
trary to  the  finding  of  Grijns  (19)  that  the  anti-beriberi  vitamin  is  les- 
sened .in  amount  as  germination  takes  place.  Mongo  is  relatively 
poor  in  vitamin  C. 

Togi  when  fresh  is  relatively  rich  in  vitamin  C;  but  after  it  is  pre- 
pared for  culinary  use,  the  vitamin  C  is  destroyed. 

The  observation  of  several  investigators  that  vitamin  C  is  increased 
when  peas,  lentils  and  beans  are  germinated,  has  been  verified  in  the 


PLANT   SOURCES  OF  VITAMINS   B   AND   C  333 

case  of  mongo.  Ten  grams  of  mongo  as  daily  supplement  to  the  scor- 
butic diet  failed  to  protect  guinea  pigs  from  scurvy,  while  5  grams  of 
fresh  togi  as  supplement  to  the  same  scorbutic  diet  cured  3  guinea  pigs 
of  the  disease. 

I  desire  to  express  my  hearty  thanks  to  Prof.  Lafayette  B.  Mendel 
for  suggesting  this  subject  to  me,  for  his  help  in  securing  the  materials 
investigated,  and  for  advice  and  criticism  during  the  progress  of  the 
work.  Part  of  the  expenses  was  defrayed  by  a  grant  from  the  Russell 
H.  Chittenden  Research  Fund  for  Physiological  Chemistry. 

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(25)  JAFFA  AND  ALBRO:  Ann.  Rept.  Cal.  Arocado  Assoc.,  1917,  85. 

(26)  JOHNS,  FINKS  AND  PAUL:  Journ.  Biol.  Chem.,  1919,  xxxvii,  497. 

(27)  MCCOLLUM:  The  Harvey  Lectures,  1916-07,  xii,  151. 

(28)  MCCOLLUM:  Journ.  Amer.  Med.  Assoc.,  1917,  Ixviii,  1379. 

(29)  MCCOLLUM,  SIMMONDS  AND  PARSONS:  Journ.  Biol.  Chem.,  1918,  xxxvi,  197. 

(30)  OSBORNE  AND  MENDEL:  Journ.  Biol.  Chem.,  1919,  37,  187. 

(31)  OSBORNE  AND  MENDEL:  Journ.  Biol.  Chem.,  1919,  xxxix,  29. 

(32)  OSBORXE  AND  MENDEL:  Journ.  Biol.  Chem.,  1920,  xxxxi,  451. 


334  FRANCISCO   O.   SANTOS 

(33)  OSBORNE  AND  MENDEL:  Journ.  Biol.  Chem.,  1920,  xxxxii,  465. 

(34)  SUGIURA  AND  BENEDICT:  Journ.  Biol.  Chem.,  1918,  xxxvi,  171. 

(35)  WHIPPLE:  Journ.  Biol.  Chem.,  1920,  xxxxiv,  175. 

(36)  WILTSHIRE:  Lancet,  1918,  (2),  811. 

Key  to  tables 

x  =  feeding  of  source  of  vitamin  B  as  daily  supplement  was  begun. 
y  =  i\  gram  yeast  replaced  other  vitamin  supplement. 

?  =  intake  of  standard  diet  could  not  be  measured  because  the  animal  scat- 
tered the  food. 

The  weekly  intake  of  standard  diet  does  not  include  the  vitamin  supplement. 

Key  to  charts 

S  =  external  sign  of  scurvy  was  first  detected. 

C  =  animal  was  chloroformed. 

•  =  }\  gram  yeast  replaced  other  vitamin  supplement. 

=  growth  on  standard  diet  alone. 

— —  =  growth  on  standard  diet  plus  vitamin  supplement  other  than 

yeast. 

. =  growth  on  standard  diet  plus  yeast. 

The  arrow,  unless  labeled  otherwise,  indicates  the  point  where  the  feeding 
of  source  of  vitamin  B  or  C  was  begun. 

The  weekly  intake  of  standard  diet  is  represented  in  graphic  form  under  each 
curve  of  weekly  body  weight. 


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